A three-step PCR protocol for construction of chimeric proteins.
A general method is described for creating chimeric proteins by transposition of subdomain-sized gene fragments. The method uses three sequential PCR steps to circumvent several limitations inherent in transposition of small insertions and has been optimized to avoid purification of intermediate products. The sequence-independence of the method permits flexibility in the choice of host molecule and insertion site.
Grandori
R
R
Department of Chemistry, Princeton University, NJ 08544, USA.
Struck
K
K
Giovanielli
K
K
Carey
J
J
D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D029968:Escherichia coli Proteins; D008565:Membrane Proteins / Q000235:genetics; D000072236:Methyl-Accepting Chemotaxis Proteins; D009841:Oligonucleotides / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D012097:Repressor Proteins
D001426:Bacterial Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; D009841:Oligonucleotides; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C081759:WrbA protein, E coli